Expression Of GAD、GABA-T、GAT-1in Rats With PMS Liver-qi Invasion Before And After Drug Intervention

Premenstrual syndrome (PMS) refers to the group of fertile women withperiodic physiology and psychological pathological changes in the lutealphase of physiological cycle which can be divided into liver-qi invasionand liver-qi depression on basis of traditional chinese medicine theorywith Liver-qi invasion as the main pathological type. Modern researchshows that the main excitability neurotransmitter Glutamic acid (Glu) andthe main inhibitive one gamma-aminobutyric acid (GABA) are both relatedto the onset and progress of liver-qi depression. Glutamate hydroxy enzyme(GAD), aminobutyric acid transaminase (GABA-T), and aminobutyric acidtransport body-1(GAT-1) are three kinds of key regulation enzyme in Glu-GABA metabolic pathways, through which Glu and GABA transformates toeach other and complete Glu-GABA metabolic pathways.This research focuseson the changes of Glu-GABA etabolic pathways in Liver-qi invasion ratsmodels in order to provide a basis for the pathogenesis of PMS Liver-qiinvasion, as well as the treatment mechanism of PMS Liver-qi invasiondrugs.ObjectiveTo explore the disease mechanism and drug intervention mechanism forPMS Liver-qi invasion.Methods1PMS liver-qi invasion rat model preparation.96rats were chosen into experiment with similar macroscopicbehavior indications and regular oestrous cycle examined by the vaginalsmear microscopy method and macroscopic behavior which were randomlydivided into four groups: control group, model group, prozac group andBXD groups. Model group rats were with conventional electricalstimulation for preparation of PMS liver-qi invasion model, while prozacrats were given prozac on the basis of electrical stimulation, and BXDgroup rats were given BaiXiangDan lavage on the basis of electricalstimulation. Control group rats were normal feed and given isodosedistilled water. All rats were killed and separation of the brainorgnizations of left hippocampal, right hippocampus, hypothalamus, andfrontal cortex which were stored in-70℃ultra-low temperature freezerafter model preparation.2microscopic index testUse High Performance Liquid Chromatography (HPLC) to determine thebrain regions Glu and GABA content; Use trizol to obtain total RNA fromthe brain area, and determine mRNA expression of GAD65, GAD67, GABA-T,and GAT–1with conventional RT-PCR method; Use the BCA kit to obtain total protein from the brain area, and determine protein expression ofGAD65, GAD67, GABA-T, and GAT–1with conventional Western blot method.Results1macroscopic behavior evaluation resultBefore modeling, there were no significant differences in the initialweight and the adaptive weight in the four groups. Compared to Controlgroup after modeling, Model rats were significantly reduced in weight(P<0.05) while significantly increased in field score (P<0.05). Comparedto Model group after modeling, both BXD group and Prozac group weresignificantly increased in weight (P<0.05) while BXD group wassignificantly reduced in field score (P<0.05).2microscopic index measurement results(1) Glu and GABA brain contentCompared to Control group, Model group was significantly higher inGlu content (P<0.05) while significantly lower in GABA content (P<0.05);Compared to Model group, Prozac group was significantly higher in bothGlu content and GABA content (P<0.05) while BXD group was significantlylower in Glu content and significantly higher in GABA content (P <0.05).(2) GAD65/67, GABA-T, GAT-1mRNA expressionCompared to Control group, Model group was significantly lower inGAD65/67mRNA expression (P<0.05) while significantly higher in GABA-T,GAT-1mRNA expression (P<0.05); Compared to Montrol group, both BXD groupand Prozac group were significantly higher in GAD65/67mRNA expression(P<0.05) while significantly lower in GABA-T, GAT-1mRNA expression(P<0.05).(3) GAD65/67, GABA-T, GAT-1protein expressionCompared to Control group, Model group was significantly lower inGAD65/67protein expression (P<0.05) while significantly higher in GABA-T,GAT-1protein expression (P<0.05); Compared to Montrol group, both BXD group and Prozac group were significantly higher in GAD65/67proteinexpression (P<0.05) while significantly lower in GABA-T, GAT-1proteinexpression (P<0.05).Conclusion1BaiXiangDan and prozac can both improve liver-qi invasion rats’ weightwhich shows curative effect. Besides, BaiXiangDan also has remarkableeffect of stabilizing liver-qi invasion rats’ emotion desease.2PMS liver-qi invasion’ pathogenesis may be associated with with a illproportion of Glu and GABA in the brain regions, which has littleassociation with the single content of Glu or GABA.3. It is maybe important links to PMS liver-qi invasion pathogenesis thatdown-regulation of GAD65, GAD67expression and up-regulation of GABA-Tand GAT-1expression subducts GABA activity throug which BaiXiangDan andprozac play a important role in treatment.4There is difference in the way for prozac and BaiXiangDan to play theeffect. Prozac may play a activated sample effect to Glu-GABA metabolicpathways through increasing irritability neurotransmitter (Glu) content,On the basis of which GAD65and GAD67expression up regulated to promptingGABA synthesis; at the same time, GAT-1and GABA-T expression are bothdown regulated to reduce GABA metabolization and uptake, through whichcomplete GABA/Glu proportional control action synergistically. AndBaiXiangDan may be raised GABA content directly through up regulate GAD65and GAD67expression and cut GAT-1and GABA-T expression through whichplaying the GABA/Glu proportional control effect.